阳和汤对人乳腺癌MCF-7细胞及其PI3K/Akt信号通路的影响
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摘要
目的:通过阳和汤含药血清干预人乳腺癌MCF-7细胞,探索阳和汤对MCF-7细胞及其内PI3K/Akt信号通路的影响。方法:制备阳和汤大鼠含药血清。MCF-7细胞设空白组,阳和汤低、高剂量组,阳性药组,阴性组,阳和汤等干预24小时、48小时、72小时,CCK-8法检测各组OD值,计算各组细胞抑制率。设空白组,阳和汤低、高剂量组,阳性药组,阳和汤等干预24小时,流式细胞术检测各组细胞凋亡的情况; WB检测各组细胞PI3K、Akt、PTEN的表达情况。结果:干预24小时、48小时、72小时后,相对于空白组,阳和汤低、高剂量组和阳性药组细胞抑制率均增高(P<0. 05)。干预24小时后,相对于空白组,阳和汤低、高剂量组和阳性药组细胞凋亡率均增高(P<0. 05)。干预24小时后,与空白组相比,阳和汤低、高剂量组和阳性药组Akt磷酸化率和PI3K磷酸化率均降低(P<0. 05或0. 01); PTEN表达量均增高(P<0. 01)。结论:阳和汤可治疗乳腺癌,其作用机制可能是通过抑制PI3K/Akt信号通路的活化,从而抑制乳腺癌细胞的增殖、诱导其凋亡,发挥抗癌作用。
Objiective: Yanghe Decoction drug-containing serum was used to intervene human breast cancer MCF-7 cells to explore the effect of Yanghe Decoction on MCF-7 cells and PI3 K/Akt signaling pathway. Methods: Yanghe Decoction rat drugcontaining serum was prepared. Blank group,Yanghe Decoction low,high dose group,positive drug group,negative group were set. After the intervention of Yanghe Decoction and other for 24 h,48 h and 72 h,the OD value of each group was detected by CCK-8 method,and the inhibition rate of each group was calculated. The blank group,the Yanghe Decoction low and high dose groups,and the positive drug group were set. After 24 h intervention,flow cytometry was used to detect the apoptosis of each group; WB was used to detect the expression of PI3 K,Akt and PTEN in each group. Results: Compared with the blank group,the inhibition rates of the Yanghe Decoction low-dose,high-dose group,and the positive drug group were all increased( P < 0.05). After 24 hours,compared with the blank group,the apoptosis rate of the Yanghe Decoction low-dose,high-dose group and the positive drug group increased( P < 0. 05). After 24 h,compared with the blank group,Akt phosphorylation rate and PI3 K phosphorylation rate were decreased in the low and high dose groups and the positive drug groups( P < 0. 05 or 0. 01);PTEN expression levels were increased( P < 0. 01). Conclusion: Yanghe Decoction can treat breast cancer. Its mechanism may be through inhibiting the activation of PI3 K/Akt signaling pathway,inhibiting the proliferation of breast cancer cells,inducing apoptosis,and exerting anti-cancer effects.
Objiective: Yanghe Decoction drug-containing serum was used to intervene human breast cancer MCF-7 cells to explore the effect of Yanghe Decoction on MCF-7 cells and PI3 K/Akt signaling pathway. Methods: Yanghe Decoction rat drugcontaining serum was prepared. Blank group,Yanghe Decoction low,high dose group,positive drug group,negative group were set. After the intervention of Yanghe Decoction and other for 24 h,48 h and 72 h,the OD value of each group was detected by CCK-8 method,and the inhibition rate of each group was calculated. The blank group,the Yanghe Decoction low and high dose groups,and the positive drug group were set. After 24 h intervention,flow cytometry was used to detect the apoptosis of each group; WB was used to detect the expression of PI3 K,Akt and PTEN in each group. Results: Compared with the blank group,the inhibition rates of the Yanghe Decoction low-dose,high-dose group,and the positive drug group were all increased( P < 0.05). After 24 hours,compared with the blank group,the apoptosis rate of the Yanghe Decoction low-dose,high-dose group and the positive drug group increased( P < 0. 05). After 24 h,compared with the blank group,Akt phosphorylation rate and PI3 K phosphorylation rate were decreased in the low and high dose groups and the positive drug groups( P < 0. 05 or 0. 01);PTEN expression levels were increased( P < 0. 01). Conclusion: Yanghe Decoction can treat breast cancer. Its mechanism may be through inhibiting the activation of PI3 K/Akt signaling pathway,inhibiting the proliferation of breast cancer cells,inducing apoptosis,and exerting anti-cancer effects.
引文
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[15] Li B,Lu Y,Wang H,et al. mi R-221/222 enhance the tu-morigenicity of human breast cancer stem cells via modulation of PTEN/Aktpathway[J]. Biomedicine&Pharmacotherapy,2016,79:93~101
[2]张英,侯炜,林洪生.中医药治疗恶性肿瘤临床研究成果与思考[J].中医杂志,2014,55(06):523~525
[3]窦建卫,郝云,杨亦奇,等.阳和汤治疗乳腺癌理论发微[J].世界中医药,2015,10(2):196~198
[4] Zhang H,Xu H L,Wang Y C,et al. 20(S)-Protopanax-adiol-Induced Apoptosis in MCF-7Breast Cancer Cell Line through the In-hibition of PI3K/AKT/m TOR Signaling Pathway[J]. International Journalof Molecular Sciences,2018,19(4):1053
[5]王维德,胡晓峰.外科证治全生集[M],2006
[6]孟李,王宁生.含药血清的制备方法研究[J].中药新药与临床药理,1999,10(5):290~292
[7]张君涛,王平,刘爱峰,等.中药含药血清制备方法的研究概述[J].中华中医药杂志,2015,30(11):4006~4009
[8]田君.阳和汤治疗乳腺癌的可行性实验研究[D].河北医科大学,2008
[9]刘亚娴,田君.阳和汤含药血清对MCF-7细胞和MDA-MB-231细胞增殖抑制作用的实验研究[J].中华中医药学刊,2009,27(2):243~246
[10]窦建卫,黄芊,赵天一,等.阳和汤对乳腺癌MDA-MB-231细胞NF-κB、IL-8表达的影响[J].四川中医,2017,10(2):44~47
[11]高永翔,沈欣,宗桂珍,等.阳和汤对裸鼠人癌移植瘤的抗肿瘤作用研究[J].中国实验方剂学杂志,2006,12(7):57~59
[12]窦建卫,任翠翠,郝云,等.阳和汤对裸鼠荷人乳腺癌组织中CD90表达的影响及其抑瘤作用[J].世界中医药,2015,10(3):391~393
[13]窦建卫,黄芊,赵天一,等.阳和汤对三阴性乳腺癌裸鼠移植瘤生长及IL-6、MMP-9表达的影响[J].中国现代医药杂志,2016,18(12):1~4
[14]田莎,王云丹,黄立中,等.阳和汤对裸鼠移植性乳腺癌骨转移模型作用机制的探讨[J].湖南中医药大学学报,2012,32(7):27~30
[15] Li B,Lu Y,Wang H,et al. mi R-221/222 enhance the tu-morigenicity of human breast cancer stem cells via modulation of PTEN/Aktpathway[J]. Biomedicine&Pharmacotherapy,2016,79:93~101